Cleavage of human serum immunoglobulin G by an immobilized pepsin preparation.
In order to obtain an efficacious and safe immunoglobulin G (IgG) preparation for intravenous use, the digestion of IgG with an immobilized pepsin (EC 3.4.23.1) preparation was studied. Thus, pepsin was immobilized onto glutaraldehyde-activated AH-Sepharose 4B under acidic conditions. THe enzymatic properties, such as proteolytic activity, pH-activity profile and heat stability, of the immobilized pepsin preparation were examined. The immobilized pepsin retained more than 40% of its proteolytic activity toward N-acetyl-L-phenylalanyl-L-3,5-diiodo-tyrosine and more than 30% toward IgG, and also remarkable stability as compared with free pepsin. The immobilized pepsin thus prepared was efficiently used for the limited cleavage of IgG and the gel-filtration effect of the column made it easily possible to yield the F(ab')2-rich fraction for intravenous use.
D002851:Chromatography, High Pressure Liquid; D004151:Dipeptides / Q000378:metabolism; D004355:Drug Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005976:Glutaral; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000378:metabolism*; D007700:Kinetics; D010434:Pepsin A / Q000378:metabolism*; D012685:Sepharose / Q000031:analogs & derivatives
D004151:Dipeptides; D004800:Enzymes, Immobilized; D007074:Immunoglobulin G; C004980:N-acetylphenylalanyl-3,5-diiodotyrosine; C024903:aminohexyl-sepharose; D012685:Sepharose; D010434:Pepsin A; D005976:Glutaral